Macrolides containing pharmaceutical compositions

ABSTRACT

A pharmaceutical compound comprising a compound of formula I and a pharmaceutically acceptable 2-amino-1,3-propanediol beside one or more pharmaceutically acceptable excipient(s).

[0001] The present invention relates to pharmaceutical compositions,e.g. for the treatment of pathological conditions comprisingco-administration of pharmaceutically active agents. Treatment includesprevention and/or therapy.

[0002] In one aspect the present invention provides a pharmaceuticallyactive compound of formula

[0003] such as disclosed in EP 427 680 (33-epi-33-chloro-FR 520 ofexample 66a), and a 2-amino-1,3-propanediol beside one or morepharmaceutically acceptable excipient(s).

[0004] Pharmaceutically acceptable excipient(s) e.g. includespharmaceutically acceptable auxiliaries, carrier(s), diluent(s). A2-amino-1,3-propanediol may comprise one or more2-amino-1,3-propanediols.

[0005] A compound of formulal I, is known to be active for the treatmentof various disorders/diseases such as e.g. inflammatory conditions,immunologically-mediated disorders, or autoimmune diseases, e.g.vasculitides, glomerulonephritides, atopic dermatitis,allergies (such asallergic contact eczema, asthma), psoriasis, systemic lupuserythematodes, rheumatoid arthritis, inflammatory bowel diseases (e.g.Crohn's disease, ulcerative colitis), multiple sclerosis,insulin-dependent diabetes, Sjögren's syndrome, endogenous posterioruveitides (in particular Behcet's disease), Hashimoto's thyroititis andto prevent rejections of xenografts or allografts, e.g. including heart,renal, hepatic or bone marrow transplants; graft vessel diseases orgraft vs host diseases.

[0006] A 2-amino-1,3-propanediol as referred to herein includes acompound of formula

[0007] wherein

[0008] R₁ is an optionally substituted straight- or branched(C₁₂₋₂₂)carbon chain, e.g. an alkyl chain, which is optionallyinterrupted by optionally substituted phenylene, and, independently ofeach other, R₂, R₃, R₄ and R₅ are H or lower alky. When the carbon chainin the meaning of R₁ is substituted, it is preferably substituted byhalogen, nitro, amino, hydroxy or carboxy. When the carbon chain isinterrupted by optionally substituted phenyl, the carbon chain ispreferably unsubstituted. When the phenylene moiety is substituted, itis preferably substituted by halogen, nitro, amino, methoxy, hydroxy orcarboxy. “Lower alkyl” includes (C₁₋₄)alkyl. Such compounds are e.g.disclosed in EP627406, EP778263, EP1002792 or WO02/06268, the relevantdisclosure of which, in particular with respect to the compounds, isincorporated herein by reference.

[0009] Preferred compounds include compounds of formula II, wherein R₁is a straight or branched, preferably straight, (C₁₃₋₂₀)carbon chainoptionally substituted by nitro, halogen, amino, hydroxy or carboxy,and, more preferably compounds of formula II, wherein R₁ is phenylalkyl,optionally substituted by halogen, e.g. wherein alkyl is (C₁₋₆)alkyl,optionally substituted by hydroxy, and wherein phenyl is substituted bystraight or branched (C₆₋₁₄)alkyl and optionally by halogen. Morepreferably, R₁ is phenyl(C₁₋₆)alkyl, wherein the phenyl group issubstituted by straight or branched, preferably straight, (C₆₋₁₄)alkyl,such as (C₆₋₁₄)alkyl-phenyl-(C₁₋₆)alkyl. If R₁ is phenylalkyl, whereinphenyl is substituted by straight or branched (C₆₋₁₄)alkyl,the phenylgroup may be substituted by alkyl in ortho, meta or para position,preferably in para position. Preferably each of R₂ to R₅ is H.

[0010] A pharmaceutical composition may comprise a compound of formula Iand a 2-amino-propanol, e.g. one as described in EP 778263, beside oneor more pharmaceutically acceptable excipient(s).

[0011] In another preferred embodiment a 2-amino-1,3,-propanediol or a2-amino-propanol is a compound having lymphocyte homing properties. Suchproperties may be identified according to a test like e.g. thefollowing: A compound to be tested or the vehicle is administered orallyby gavage to rats. Tail blood for hematological monitoring is obtainedon day 11 to give the baseline individual values, and at 2, 6, 24, 48and 72 hours after application. A lymphocyte homing agent is a compoundwhich depletes peripheral blood lymphocytes by more than 50% six hours,e.g. after administration of a dose smaller than 5 mg/kg, preferablysmaller than 3 mg/kg.

[0012] A still more preferred compound of a compound of formula II is acompound of formula

[0013] e.g. 2-amino-2[2-(4-octylphenyl)ethyl]propane-1,3-diolhydrochloride. A 2-amino-1,3-propanediol, e.g. a compound of formula II,may be useful, on the basis of observed activity, e.g. as described inEP 627406, as immunomodulators, e.g. immunosuppressants, e.g. in thetreatment of allograft rejections. E.g. as described in WO 98/22100,2-amino-1,3-propanediols, e.g. of formula II, may inhibit graft vesseldisease and are particularly indicated to prevent or treat chronicrejection in a transplanted organ, and additionally may suppressxenograft rejection.

[0014] According to the present invention a 2-amino-1,3-propanediol or a2-amino-propanol is pharmaceutically active and pharmaceuticallyacceptable.

[0015] A compound of formula I and a 2-amino-1,3-propanediol includecompounds in any form, e.g. in free form, in the form of a salt, in theform of a solvate and in the form of a salt and a solvate, where suchforms exist. The compounds according to the present invention

[0016] in free form may be converted into a corresponding compound inthe form of a salt;

[0017] in free form or in the form of a salt and in the form of asolvate may be converted into a corresponding compound in free form orin the form of a salt in unsolvated form;

[0018] and vice versa.

[0019] The pharmaceutical activity of a compound in free form is in asimilar range as that of a compound in salt/solvate form. A solvateincludes a hydrate. A salt includes a pharmaceutically acceptable salt.

[0020] E.g. pharmaceutically acceptable salts of a compound of formulaII include salts of a compound of formula II with an acid, e.g.including an inorganic acid, such as hydrochloric acid, hydrobromic acidand sulphuric acid, and including an organic acid, such as acetic acid,fumaric acid, maleic acid, benzoic acid, citric acid, malic acid,methanesulfonic acid and benzenesulfonic acid, and, if a carboxy groupis present, salts with

[0021] metals such as sodium, potassium, calcium and aluminium,

[0022] amines, such as triethylamine, and

[0023] dibasic amino acids, such as lysine.

[0024] In a preferred embodiment the compounds of formula II or those asdescribed in EP1002792 includes phosphates of said compounds.

[0025] A compound of formula I and a compound of formula II may exist inisomeric forms and the present invention includes a compound of formulaI and a 2-amino-1,3-propanediol according to the present invention inany isomeric form and any isomeric mixture. E.g. if a compound offormula I and a 2-amino-1,3-propanediol according to the presentinvention has one or more asymmetric centers in the molecule, thepresent invention includes the compound in the form of various opticalisomers, as well as racemates, diastereoisomers and mixtures thereof.

[0026] We have now surprisingly found, that in an animal model ofexperimental autoimmune uveititis (EAU) wherein a compound of formula Ior a 2-amino-1,3-propanediol show pharmaceutical activity, an increasedactivity is achieved when the compound of formula I and the2-amino-1,3-propanediol, e.g. a compound of formula II, areco-administered in e.g. suboptimal doses. That increased activity of theco-administered compounds is remarkable higher than that of each singlecompound tested at the e.g. suboptimal dose level. The same principle isbelieved to be valid for all diseases wherein either a compound offormula I or a 2-amino-1,3-propanediol shows pharmaceutical activity,e.g. including inflammatory conditions, immunologically-mediateddisorders, or autoimmune diseases, e.g. vasculitides,glomerulonephritides, atopic dermatitis,allergies (such as allergiccontact eczema, asthma), psoriasis, systemic lupus erythematodes,rheumatoid arthritis, inflammatory bowel diseases (e.g. Crohn's disease,ulcerative colitis), multiple sclerosis, insulin-dependent diabetes,Sjögren's syndrome, endogenous posterior uveitides (in particularBehcet's disease), Hashimoto's thyroititis and to prevent rejections ofxenografts or allografts, e.g. including heart, renal, hepatic or bonemarrow transplants; graft vessel diseases or graft vs host diseases.

[0027] In another aspect the present invention provides a packagecomprising a compound of formula I in the form of a pharmaceuticalcomposition beside one or more pharmaceutically acceptable excipient(s)and comprising instructions for simultaneous or sequentialadministration of a 2-amino-1,3-propanediol.

[0028] In another aspect the present invention provides a packagecomprising a 2-amino-1,3-propanediol in the form of a pharmaceuticalcomposition beside one or more pharmaceutically acceptable excipient(s)and comprising instructions for simultaneous or sequentialadministration of a compound of formula I.

[0029] In another aspect the present invention provides a pharmaceuticalkit, e.g. a package, comprising a compound of formula I in the form of apharmaceutical composition beside one or more pharmaceuticallyacceptable excipient(s), and a 2-amino-1,3-propanediol in the form of apharmaceutical composition beside one or more pharmaceuticallyacceptable excipient(s) in the same package.

[0030] In another aspect the present invention provides a method ofimproving the pharmaceutically activity of a compound of formula I asdescribed earlier which method comprises co-administrating a compound offormula I and a 2-amino-1,3-propanediol to a subject in need of atreatment with a compound of formula I and/or with a2-amino-1,3-propanediol.

[0031] The compound of formula I and a 2-amino-1,3-propanediol may beco-administrated in different ways:

[0032] a) In the form of fixed combinations, comprising a compound offormula I and a pharmaceutically active 2-amino-1,3-propanediol in thesame pharmaceutical composition;

[0033] b) In the form of a (pharmaceutical) kit, in which a compound offormula I and a 2-amino-1,3-propanediol are present in the form ofseparate pharmaceutical compositions, sold in the same package, e.g.with instruction for co-administration;

[0034] c) In the form of free combinations, in which a compound offormula I and a 2-amino-1,3-propanediol are packaged separately, e.g. inthe form of pharmaceutical compositions, wherein each of the packagesinclude instructions for simultaneous or sequential administration.

[0035] The most efficient ratio of a compound of formula I and a2-amino-1,3-propanediol may be dependent e.g. on the indication to betreated. Appropriate dosages and dosage ranges will of course varydepending upon, for example, the active compounds of the presentinvention used, the host, the mode of administration and the nature andseverity of the conditions being treated. However, in general, forsatisfactory results in larger mammals, for example humans, an indicateddaily dosage is in the range of that which is known for a macrolactamlike a pharmaceutically active compound of formula I and apharmaceutically active 2-amino-1,3-propanediol, e.g. below theseoptimal dosages, administered, for example, in divided doses, e.g. up tofour times a day. In general, satisfactory results are indicated to beobtained systemically at daily dosages of from about 0.5 mg/kg to about15 mg/kg, preferably 1 mg/kg to about 15 mg/kg animal/human body weightof a compound of formula I and form about 0.005 mg/kg to 0.1 mg/kg,prfereably 0.01 mg/kg to 0.1 mg/kg of a compound of formula II.Preferred ratios of compound I to II are in a range between about 3000to 10, preferably 500 to 10.

[0036] The active compounds of the present invention may be administeredby any conventional route, e.g. systemically, for example orally, e.g.in form of tablets or capsules, or parenterally, e.g. in the form ofinjectable solutions or suspensions; and topically, such asepicutaneous, intranasal, intratracheal administration.

[0037] In another aspect the present invention provides a method oftreatment of diseases, wherein a compound of formula I and/or a2-amino-1,3-propanediol are pharmaceutically active, e.g. includinginflammatory conditions, immunologically-mediated disorders, orautoimmune diseases, e.g. vasculitides, glomerulonephritides, atopicdermatitis,allergies (such as allergic contact eczema, asthma),psoriasis, systemic lupus erythematodes, rheumatoid arthritis,inflammatory bowel diseases (e.g. Crohn's disease, ulcerative colitis),multiple sclerosis, insulin-dependent diabetes, Sjögren's syndrome,endogenous posterior uveitides (in particular Behcet's disease),Hashimoto's thyroititis and to prevent rejections of xenografts orallografts, e.g. including heart, renal, hepatic or bone marrowtransplants; graft vessel dieseases or graft vs host diseases,comprising administering to a subject in need of such treatment aneffective amount of a compound of formula I and a2-amino-1,3-propanediol; e.g. in the form of

[0038] a fixed combination, e.g. in the form of a pharmaceuticalcomposition,

[0039] a kit, e.g. in the form of a pharmaceutical composition of acompound of formula I and a pharmaceutical composition of a2-amino-1,3-propanediol in the same package,

[0040] a package comprising a compound of formula I or a2-amino-1,3-propanediol, e.g. in the form of pharmaceuticalcompositions, and comprising instructions for simultaneous or sequentialco-administration.

[0041] In a preferred embodiment of the present invention a compound offormula I is co-administered with a compound of formula IIa.

[0042] The compound of formula I and a 2-amino-1,3-propanediol may beadministered as the sole ingredients or together with other drugs inimmunomodulating regimens or other anti-inflammatory agents. For examplethe compounds may be used in combinations with cyclosporins, rapamycinsor other ascomycins, or their immunosuppressive analogs, e.g.cyclosporin A, cyclosporin G, FK-506, rapamycin,40-O-(2-hydroxy)ethyl-rapamycin, etc.; corticosteroids;cyclophosphamide; azathioprene; methotrexate; brequinar; leflunomide;mizoribine; mycophenolic acid; mycophenolate mofetil;15-deoxyspergualine; immunosuppressive monoclonal antibodies, e.g.monoclonal antibodies to leukocyte receptors, e.g. to MHC, CD2, CD3,CD4, CD7, CD25, CD28, B7, CD40, CD45 or CD58 or to their ligands; orother immunomodulatory compounds, e.g. CTLA4-Ig.

[0043] Tests for the determination of the activity of a compound offormula I and 2-amino-1,3-propanediols are known. A model ofexperimental autoimmune uveitits (EAU) as described e.g. in Clin.Immunol. Immunopathol. 1986; 39: 329-336, McAllister et al.(1986) can beused.

[0044] In another aspect the present invention provides the use of acompound of formula I together with a 2-amino-1,3-propanediol, e.g.administered either simultanously or sequentially, as a pharmaceutical;and

[0045] the use of a combination of a compound of formula I and a2-amino-1,3-propanediol in the preparation of a medicament for thetreatment of diseases, wherein the compound of formula I or a2-amino-1,3-propanediol are pharmaceutically active. Treatment includesprevention and/or therapy.

DESCRIPTION OF THE FIGURES

[0046] In FIG. 1, results of the Experimental autoimmune uveitits (EAU)test in rats are indicated. The rats were treated orally, once daily,for 14 days, either with

[0047] ♦: Control (placebo, i.e. drug vehicle/water),

[0048] ▪: Compound of formula IIa (0.1 mg),

[0049] ▴: Compound of formula I (15 mg), or

[0050]

: Compound of formula IIa (0.1 mg)+Compound of formula I (15 mg).

[0051] The weights in mg are indicated in mg/kg body weight/day. Themeans of scores of both eye lesions were determined as described in theexamples within 20 days after immunization.

[0052] In FIG. 2, results of the Experimental autoimmune uveitits (EAU)test in rats are indicated. The rats were treated orally, once daily,for 14 days, either with

[0053] ♦: Control (placebo, i.e. drug vehicle/water),

[0054] ▪: Compound of formula IIa (0.1 mg),

[0055] ▴: Compound of formula I (15 mg),

[0056]

: Compound of formula I (15 mg)+Compound of formula IIa (0.1 mg), 14days, or

[0057] *: Compound of formula I (15 mg)+Compound of formula IIa (0.1mg), 28 days.

[0058] Other parameters are as described for FIG. 1.

EXAMPLES Example 1 EAU Test System Experimental Autoimmune Uveitits(EAU)

[0059] The model of EAU used, is similar to that described previously byMcAllister et al.(1986) (McAllister C G, Vistica B P, Sekura R, KuwabaraT, Gery I. The effects of pertussis toxin on the induction and transferof experimental autoimmune uveoretinitis. Clin Immunol Immunopathol1986; 39: 329-336). The animals (5 rats per group) are injected underether anesthesia into the right foodpat with 50 μg of purified bovineretinal S-antigen and with 1 μg pertussis toxin (Difco) intraperitoneally on day 1. The antigen is diluted with phosphate-bufferedsaline and mixed 1:1 (v/v) with Freund's complete adjuvant and Bacto MTuberculosis H37 RA (Difco). The volume injected is 0.1 ml, containing50 μl complete adjuvant and 1.14 mg of Mycobacterium tuberculosis. Thisprocedure induces a fulimant disease in all animals which is observedearliest 9-10 days after immunisation. The eye lesions in untreatedanimals develop in allmost all animals to severity grade 4 (see section“Evaluation of EAU”).

Treatment Dosages

[0060] The animals are treated by gavage with daily dosages of 15 mg/kgof a compound of formula I or 0.1 mg/kg of a compound of formula IIaalone; or with a combination of both compounds at the same dosagelevels. The treatment is started 2 hours before immunisation andperformed once daily on 14 consecutive days. Control animals are treatedsimilarly with placebo/water alone (placebo: drug vehicle alone).

Evaluation of EAU

[0061] Starting on day 7 after immunisation the animals are examinedwith an ophtalmoscop (Heine, Beta 200) for inflammatory changes daily upto day 20. The extent of ocular inflammation is semi-quantitativelyassessed with scores from 0 to 4 (for 1 eye).

[0062] 0: normal;

[0063] 1: iris hyperemia;

[0064] 2: iris hyperemia with vasculare dilatation;

[0065] 3: early fibrinous exudate in the anterior chamber and moderateiris cell infiltration; and

[0066] 4: large fibrin clot in anterior chamber or fibrin plugging ofthe pupil and severe iris cell infiltration.

Results

[0067] Animals (LEWIS rats) are treated orally either with 0.1 mg/kg ofa compound of formula IIa or 15 mg/kg/day of a compound of formula I arenot distinctively different from placebo controls in the course andintensity of the disease (see e.g. TABLE 1 and FIG. 1). In contrast, theadministration of both compounds at the same dosage levels causes aremarkably delay in onset of the disease (6 days later than in controls)and the intensity of the inflammation is remarkably less. The highestmean score of treated animals is 4.4 on day 19 compared with 8.0 on day11 in controls. TABLE 1 EAU SCORE MAX Test group Dosis POS EAU 10 EAU1^(st) SCORE DAY Controls 0.0 5/5 5/5 10.0 (0.0) 8.0 (0.0) 11 Cpd IIa0.1   4/4*⁾ 2/5 10.5 (0.6) 8.0 (0.0) 13 Cpd I 15   5/5 1/5 11.8 (1.3)6.8 (1.3) 14 Cpd IIa + 1.5 + 15 4/5 0/5   16 (1.4) 4.4 (3.3) 19 Cpd I

[0068] In TABLE 1 under “EAU POS” and “EAU 10” the number of affectedanimals/number of animals per group until end of the study (EAU POS),and on day 10 (EAU 10), respectively are indicated. The followingabbreviations are used:

[0069] Cpd I: Compound of formula I

[0070] Cpd IIa: Compound of formula IIa

[0071] Controls: Placebo (drug vehicles/water)

[0072] Dosis: oral, in mg/kg/day

[0073] EAU POS: Number of EAU positive rats

[0074] EAU 10: Incidence of EAU on day 10

[0075] EAU 1st: Day of first signs of EAU (mean±SD)

[0076] SCORE MAX: Maximum score (mean of both eye lesions±SD)

[0077] The results from the EAU-test (used as an assay example) indicatethat the combination of a suboptimal dosage of a compound of formula Iplus a suboptimal dosage of a compound of formula IIa is significantlysuperior in activity against EAU, than the use of a suboptimal dosage ofa compound of formula I, or of a suboptimal dosage of a compound offormula IIa, respectively, alone (Table 1 and FIG. 1). That effectallows the use of a suboptimal dosage of a compound of formula I plus asuboptimal dosage of a compound of formula IIa.

Example 2

[0078] Animals (6 rates per group) were treated as described in example1 with the doses given in table 2. As depicted in table 2 and FIG. 2 thecombination tretament shows superior results over the treatment withsingle compounds. From the animal treated on 14 days with bothcompounds, the first clinical signs were observed in one animal on day16. Complete suppression of symptoms was observed also during the 4-weektreatment period. Symptoms appeared earliest in one animal on day 34, 6days after the last treatment. TABLE 2 EAU SCORE MAX Test group DosisPOS EAU 10 EAU 1^(st) SCORE DAY Controls — 6/6 6/6  9.3 (0.5) 8.0 (0.0)12 Cpd IIa 0.1 6/6 0/6 11.2 (1.0) 8.0 (0.0) 14 Cpd I 15   6/6 2/6 11.5(0.8) 6.0 (2.5) 14 Cpd I + 15 + 0.1 6/6 0/6 18.0 (1.1) 8.0 (0.0) 21 CpdIIa (14×) Cpd I + 15 + 0.1 6/6 0/6 34.8 (0.4) 7.2 (1.3) 37 Cpd IIa (28×)

[0079] The meanings are as in legend to table 1

[0080] The results from the EAU-test (used as an assay example) indicatethat the development of the disease can be prevented when treatment isperformed for 4 weeks starting at immunization with a combination ofcompounds I and IIa.

1. A pharmaceutical composition comprising a compound of formula

and a 2-amino-1,3-propanediol beside one or more pharmaceuticallyacceptable excipient(s).
 2. A package comprising a compound of formula Ias defined in claim 1 in the form of a pharmaceutical composition besideone or more pharmaceutically acceptable excipient(s) and comprisinginstructions for simultaneous or sequential administration of a2-amino-1,3-propanediol.
 3. A package comprising a2-amino-1,3-propanediol in the form of a pharmaceutical compositionbeside one or more pharmaceutically acceptable excipient(s) andcomprising instructions for simultaneous or sequential administration ofa compound of formula I as defined in claim
 1. 4. A pharmaceutical kit,comprising a compound of formula I as defined in claim 1 in the form ofa pharmaceutical composition beside one or more pharmaceuticallyacceptable excipient(s) and a 2-amino-1,3-propanediol in the form of apharmaceutical composition beside one or more pharmaceuticallyacceptable excipient(s) in the same package.
 5. Use of a compound offormula I as defined in claim 1 in combination with a pharmaceuticallyactive 2-amino-1,3-propanediol as a pharmaceutical.
 6. Use of acombination of a compound of formula I as defined in claim 1 and a2-amino-1,3-propanediol in the preparation of a medicament for thetreatment of diseases, wherein a compound of formula I and a2-amino-1,3,-propanediol are pharmaceutically active.
 7. A method ofimproving the pharmaceutical activity of a compound of formula I and ofa pharmaceutically acceptable 2-amino-1,3-propanediol which methodcomprises co-administrating a compound of formula I and a2-amino-1,3-propanediol to a subject in need of a treatment with acompound of formula I and/or with a 2-amino-1,3-propanediol.
 8. A methodof treatment of diseases, wherein a compound of formula I as defined inclaim 1 and a 2-amino-1,3-propanediol are pharmaceutically active,comprising administering to a subject in need of such treatment aneffective amount of a compound of formula I as defined in claim 1 and a2-amino-1,3-propanediol.
 9. A method, a pharmaceutical composition, apharmaceutical kit, a package or the use as claimed in any one of thepreceding claims, wherein the 2-amino-1,3-propanediol is a compound offormula

wherein R₁ is an optionally substituted straight- or branched(C₁₂₋₂₂)carbon chain, and, independently of each other, R₂, R₃, R₄ andR₅ are H or lower alky.
 10. A method, a pharmaceutical composition, apharmaceutical kit, a package or the use as claimed in claim 9, whereina compound of formula II is of formula